Katalin Kocsis

Localization of putative glutamatergic/aspartatergic neurons projecting to the supraoptic nucleus area of the rat hypothalamus.

Oxytocin and vasopressin neurosecretory neurons of the supraoptic nucleus receive a rich glutamatergic innervation. The nerve cells of this prominent structure express various ionotropic and metabotropic glutamate receptor subtypes and there is converging evidence that glutamate acts as an excitatory transmitter in the control of release of oxytocin and vasopressin synthesized in this cell group. The location of the glutamatergic neurons projecting to this hypothalamic region is unknown. The aim of the present investigation was to study this question. [3H]d‐aspartate, which is selectively taken up by high‐affinity uptake sites at presynaptic endings that use glutamate as a transmitter, and is transported back to the cell body, was injected into the supraoptic nucleus area. The neurons retrogradely labelled with [3H]d‐aspartate were detected autoradiographically. Labelled nerve cells were found in several diencephalic and telencephalic structures, but not in the brainstem. Diencephalic cell groups included the supraoptic nucleus itself, its perinuclear area, hypothalamic paraventricular, suprachiasmatic, ventromedial, dorsomedial, ventral premammillary, supramammillary and thalamic paraventricular nuclei. Within the telencephalon, labelled neurons were detected in the septum, amygdala, bed nucleus of the stria terminalis and preoptic area. The findings provide neuromorphological data on the location of putative glutamatergic neurons projecting to the supraoptic nucleus and its perinuclear area. Furthermore, they indicate that local putative glutamatergic neurons as well as several diencephalic and telencephalic structures contribute to the glutamatergic innervation of the cell group and thus are involved in the control of oxytocin and vasopressin release by neurosecretory neurons of the nucleus.

Identification of CNS neurons involved in the innervation of the epididymis: A viral transneuronal tracing study

Cell groups of the spinal cord and the brain transsynaptically connected with the epididymis (caput, cauda) were identified by means of the viral transneuronal tracing technique. Pseudorabies virus was injected into the caput or the cauda epididymidis, and after survival times 4 and 5 days, the spinal cord and brain were processed immunocytochemically. Virus-labeled neurons could be detected in the preganglionic sympathetic neurons (lower thoracic and upper lumbar segments) and following virus injection into the cauda epididymidis, also in the sacral parasympathetic nucleus (L6-S1). Virus-infected perikarya were present in several brain stem nuclei (lateral reticular nucleus, gigantocellular and paragigantocellular nucleus, A5 noradrenergic cell group, caudal raphe nuclei, locus coeruleus, Barringtons nucleus, nucleus of the solitary tract, periaqueductal gray) and in the diencephalon (hypothalamic paraventricular nucleus, lateral hypothalamus). At the longer survival time, some telencephalic structures also exhibited virus-labeled neurons. The distribution of infected neurons in the brain was similar after virus injection into the caput or cauda epididymidis; however, earlier onset of infection was observed after inoculation into the cauda. The present findings provide the first morphological data on a multisynaptic circuit of neurons innervating the epididymis and presumably involved in the control of epididymal functions. reserved.

Transneuronal labelling of nerve cells in the CNS of female rat from the mammary gland by viral tracing technique

Using the viral transneuronal tracing technique, the cell groups in the CNS transneuronally connected with the female mammary gland were detected. Lactating and non-lactating female rats were infected with pseudorabies virus injected into the mammary gland. The other group of animals was subjected to virus injection into the skin of the back. Four days after virus injection, infected neurons detected by immunocytochemistry, were present in the dorsal root ganglia ipsilateral to inoculation and in the intermediolateral cell column of the spinal cord. In addition, a few labelled cells could be detected in the dorsal horn and in the central autonomic nucleus (lamina X) of the spinal cord. At this survival time several brain stem nuclei including the A5 noradrenergic cell group, the caudal raphe nuclei (raphe obscurus, raphe pallidus, raphe magnus), the A1/C1 noradrenergic and adrenergic cell group, the nucleus of the solitary tract, the area postrema, the gigantocellular reticular nucleus, and the locus coeruleus contained virus-infected neurons. In some animals, additional cell groups, among others the periaqueductal gray and the red nucleus displayed labelling. In the diencephalon, a significant number of virus-infected neurons could be detected in the hypothalamic paraventricular nucleus. In most cases, virus-labelled neurons were present also in the lateral hypothalamus, in the retrochiasmatic area, and in the anterior hypothalamus. In the telencephalon, in some animals a few virus-infected neurons could be found in the preoptic area, in the bed nucleus of the stria terminalis, in the central amygdala, and in the somatosensory cortex. At the longer (5 days) survival time each cell group mentioned displayed immunopositive neurons, and the number of infected cells increased. The pattern of labelling was similar in animals subjected to virus inoculation into the mammary gland and into the skin. The distribution and density of labelling was similar in lactating and non-lactating rats. The present findings provide the first morphological data on the localization of CNS structures connected with the preganglionic neurons of the sympathetic motor system innervating the mammary gland. It may be assumed that the structures found virus-infected belong to the neuronal circuitry involved in the control of the sympathetic motor innervation of the mammary gland.

Metabotropic glutamate receptor in vasopressin, CRF and VIP hypothalamic neurones

THE excitatory amino acid glutamate, acting via ionotropic and metabotropic glutamate receptors, appears to play an important role in the control of neuroendocrine functions. The aim of the present investigations was to determine whether hypothalamic neurones which synthesize arginin-vasopressin (AVP), CRF and VIP express metabotropic glutamate receptor (mGluR). Double-label immunocytochemistry and the mirror technique were used. We found that AVP immunoreactive neurones of the paraventricular, supraoptic and suprachiasmatic nuclei contain mGluR1a, but the number of double-labelled neurones was different in the three cell groups. mGluR1a was present in a significant number of paraventricular CRF nerve cells, and in almost all VIP neurones of the SCHN. These results support the view that the excitatory transmitter glutamate may directly influence AVP, CRF and VIP neurones of the three hypothalamic cell groups.

Metabotropic glutamate receptor in GHRH and β-endorphin neurones of the hypothalamic arcuate nucleus

GROWTH hormone-releasing hormone (GHRH) and β-endorphin are mainly synthesized in neurones of the hypothalamic arcuate nucleus. Arcuate neurones also contain both ionotropic and metabotropic glutamate receptors. The aim of present study was to investigate whether glutamate receptors are present in GHRH and β-endorphin containing nerve cells of this hypothalamic area. Using double-label immunocytochemistry as well as the mirror technique, we found that almost all GHRH and β-endorphin immunoreactive arcuate neurones contain the metabotropic glutamate receptor 1a. The observations provide morphological evidence for the view that glutamate, which appears to be a major excitatory neurotransmitter in the hypothalamus, may directly stimulate GHRH and β-endorphin neurones of the medial hypothalamus.

Origin of the chicken splenic reticular cells influences the effect of the infectious bursal disease virus on the extracellular matrix

The effects of infectious bursal disease virus (IBDV) (strain F52/70) infection were studied by immunohistochemical methods on the splenic extracellular matrix (ECM). The major fibrillar components of the ECM, the type I and type III collagens and the main ECM organizing glycoproteins (laminin, tenascin and fibronectin) were monitored up to 11 days post-infection (d.p.i.). By 3 d.p.i., the collagens that form the basic scaffold of the antigen-trapping region of the spleen are destroyed, which is followed by deterioration of the glycoproteins. The ECM in the red pulp and the other regions of the white pulp (periarteriolar lymphatic sheath and germinal centre) seem to be normal. The reason for the significantly different pathological alterations in the ECM between the two regions of the spleen may be explained by the origin of the reticular cells. The reticular cells in the antigen-trapping zone and other splenic regions are of haemopoietic and mesenchymal origins, respectively. Possibly, the reticular cells of the haemopoietic origin are more susceptible for the IBDV infection than the mesenchymal ones. Development of the antigen-trapping, B-cell-dependent zone of the splenic white pulp precedes that of the periarteriolar lymphatic sheath and germinal centre, which suggests that this region may contribute to B-cell maturation. Damage of the ECM in the antigen-trapping zones results in impairment of tissue organization, which may contribute to the permanent immunosuppression.

Distribution of PACAP and its mRNA in several nonneural tissues of rats demonstrated by sandwich enzyme immunoassay and RT-PCR technique

The presence of PACAP in various organs was previously demonstrated using immunohistochemistry and radioimmunoassay. The aim of our work was to get information whether the presence of immunoreactive PACAP in various organs, mainly in the gastric mucosa, also indicates the place of its synthesis. The immunoreactive PACAP and its mRNA were measured in parallel assays using sandwich enzyme immunoassay (S-EIA) and RT-PCR technique. PACAP and its mRNA were demonstrated in the pancreas, testes, adrenal glands, ovaries, and in the oxyntic mucosa of the stomach. These results support our previous observation that PACAP is present not only in the nervous system and endocrine glands, but might be synthetized in the oxyntic mucosa of the stomach as well.

Chicken dendritic cells and type II pneumocytes express a common intracellular epitope

1. CVI-ChNL 74·3, a dendritic cell-specific monoclonal antibody (mAb) also identifies chicken lung granular pneumocytes (type II pneumocytes), which produce surfactant. 2. The 74·3 mAb does not cross-react with any other avian or mammalian granular pneumocyte, and provides a convenient tool for monitoring the status of type II pneumocytes in the chicken lung.

Dual secretion locations on type II cells in the avian lung suggest local as well as general roles of surfactant

Transmission electron microscopy indicates that the avian lung surfactant may be secreted in two directions: a) into air passages of parabronchus, atrium and infundibulum where it forms a trilaminar substance serving the respiratory role and b) to the basolateral surface—intercellular space—of type II pneumocytes, contributing to the innate and adoptive immune responses of lung. Basolateral secretion may be confirmed by the presence of trilaminal substance in the intercellular space of type II pneumocytes. Fusion of surfactant containing vesicles with the lateral plasma membrane may result in membrane fusion of neighboring cells and subsequently formation of multinucleated giant cell. The indistinct and in some places discontinuous basal lamina in the parabronchial atrium and infundibulum permits the hydrophilic surfactant proteins to spread into the interstitium of air‐blood capillary region. The hydrophilic surfactant proteins may activate lung interstitial macrophages to migrate into the air passages where they appear as “free avian respiratory macrophages.” Therefore, in the interstitium the hydrophilic surfactant proteins are essential soluble components of innate immunity. J. Morphol. 277:1062–1071, 2016.